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Reverse Transcription Cross-Priming Amplification-Nucleic Acid Test Strip for Rapid Detection of Porcine Epidemic Diarrhea Virus

文献类型: 外文期刊

作者: Wang, Feng-Xue 1 ; Yuan, Dan-Yi 3 ; Jin, Ya-Nan 4 ; Hu, Lin 5 ; Sun, Zhi-Yong 3 ; He, Qian 3 ; Zhao, Shi-Hua 2 ; Zhan, Sh 1 ;

作者机构: 1.Chinese Acad Agr Sci, Inst Special Econ Anim & Plant Sci, State Key Lab Special Econ Anim Mol Biol, Jingyue Econ & Technol Dev Zone, 4899 Juye Ave, Changchun 130112, Jilin, Peoples R China

2.Inner Mongolia Acad Agr & Anim Husb Sci, Vet Res Inst, Hohhot 010031, Inner Mongolia, Peoples R China

3.Sino USA SiChuan Nabii Biotech Co Ltd, Chengdu 610041, Sichuan, Peoples R China

4.Ustar Biotechnol Hangzhou Ltd, Hangzhou 310012, Zhejiang, Peoples R China

5.Ustar Biotechnol

期刊名称:SCIENTIFIC REPORTS ( 影响因子:4.379; 五年影响因子:5.133 )

ISSN: 2045-2322

年卷期: 2016 年 6 卷

页码:

收录情况: SCI

摘要: Porcine epidemic diarrhea virus (PEDV) is a highly transmissible coronavirus that causes a severe enteric disease particularly in neonatal piglets. In this study, a rapid method for detecting PEDV was developed based on cross-priming amplification and nucleic acid test strip(CPA-NATS). Five primers specific for the N gene sequence of PEDV were used for the cross-priming amplification. Detection of amplification products based on labeled probe primers was conducted with strip binding antibody of labeled markers. The CPA method was evaluated and compared with a PCR method. The reverse transcription CPA system was further optimized for detecting PEDV RNA in clinical specimens. Results showed that the method was highly specific for the detection of PEDV, and had the same sensitivity as PCR, with detection limit of 10(-6) diluted plasmid containing the target gene of PEDV. It was also successfully applied to detecting PEDV in clinical specimens. The reverse transcription CPA-NATS detection system established in this study offers a specific, sensitive, rapid, and simple detection tool for screening PEDV, which can contribute to strategies in the effective control of PEDV in swine.

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